scholarly journals HEALING OF BROKEN LINEAR DICENTRIC CHROMOSOMES IN YEAST

Genetics ◽  
1984 ◽  
Vol 106 (2) ◽  
pp. 207-226
Author(s):  
James E Haber ◽  
Patricia C Thorburn
Keyword(s):  
Dna Sequences ◽  
Meiotic Recombination ◽  
Recombinant Dna ◽  
Dicentric Chromosome ◽  
Homologous Region ◽  
Repeated Dna ◽  
Circular Chromosome ◽  
Dicentric Chromosomes ◽  
Chromosome Iii ◽  
Linear Chromosomes

ABSTRACT In yeast, meiotic recombination between a linear chromosome III and a haploid-viable circular chromosome will yield a dicentric, tandemly duplicated chromosome. Spores containing apparently intact dicentric chromosomes were recovered from tetrads with three viable spores. The spore containing the dicentric inherited URA3 (part of the recombinant DNA used to join regions near the ends of the chromosome into a circle) as well as HML, HMR and MAL2 (located near the two ends of a linear but deleted from the circle). The Ura+ Mal+ colonies were highly variegated, giving rise to as many as seven distinctly different stable ("healed") derivatives, some of which were Ura+ Mal+, others Ura+ Mal- and others Ura- Mal+. The colonies were also sectored for five markers (HIS4, LEU2, CRY1, MAT and THR4) initially heterozygous in the tandemly duplicated dicentric chromosome.—Southern blot and genetic analyses have demonstrated that these stable derivatives arose from mitotic break-age of the dicentric chromosome, followed by one of several different healing events. The majority of the stable derivatives contained circular or linear chromosomes apparently resulting from homologous recombination between a broken chromosome end and a homologous region on the other end of the original dicentric duplicated chromosome. A smaller proportion of events resulted in apparently uniquely healed linear chromosomes in which the broken chromosome acquired a new telomere. In two instances we recovered chromosome III partially duplicated with a novel right end. We have also found one derivative that had also experienced rearrangement of repeated DNA sequences found adjacent to yeast telomeres.

scholarly journals MEIOTIC AND MITOTIC BEHAVIOR OF DICENTRIC CHROMOSOMES IN SACCHAROMYCES CEREVISIAE

Genetics ◽  
1984 ◽  
Vol 106 (2) ◽  
pp. 185-205
Author(s):  
James E Haber ◽  
Patricia C Thorburn ◽  
David Rogers
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Recombinant Dna ◽  
Ring Chromosome ◽  
Dicentric Chromosome ◽  
Linear Chromosome ◽  
Dicentric Chromosomes ◽  
Ring Chromosomes ◽  
Mitotic Instability ◽  
Unequal Number ◽  
Chromosome Iii

ABSTRACT Meiotic recombination between a circular and a linear chromosome in Saccharomyces cerevisiae has been investigated. The circle was a haploid-viable derivative of chromosome III constructed by joining regions near the two chromosome ends via a recombinant DNA construction: (HMR/MAT-URA3-pBR322-MAT/HML) and was also deleted for MAL2(which therefore uniquely marks a linear chromosome III). Recombination along chromosome III was measured for eight intervals spanning the entire length of the circular derivative. Only 25% of all tetrads from a ring/rod diploid contained four viable spores. These proved to be cases in which there was either no recombination along chromosome III or in which there were two-strand double crossovers or higher order crossovers that would not produce a dicentric chromosome.—At least half of the tetrads with three viable spores included one Ura+ Mal+ spore that was genetically highly unstable. The Ura+ Mal+ spore colonies gave rise to as many as seven genetically distinct, stable ("healed") derivatives, some of which had lost either URA3 or MAL2. Analysis of markers on chromosome III suggests that dicentric chromosomes frequently do not break during meiosis but are inherited intact into a haploid spore. In mitosis, however, the dicentric chromosome is frequently broken, giving rise to a variety of genetically distinct derivatives. We have also shown that dicentric ring chromosomes exhibit similar behavior: at least half the time they are not broken during meiosis but are broken and healed during mitosis.—The ring/rod diploid can also be used to determine the frequency of sister chromatid exchange (SCE) along an entire yeast ring chromosome. We estimate that an unequal number of SCE events occurs in approximately 15% of all cells undergoing meiosis. In contrast, the mitotic instability (and presumably SCE events) of a ring chromosome is low, occurring at a rate of about 1.2 x 10-3 per cell division.

Effects of excess centromeres and excess telomeres on chromosome loss rates

1991 ◽  
Vol 11 (6) ◽  
pp. 2919-2928
Author(s):  
K W Runge ◽  
R J Wellinger ◽  
V A Zakian
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Division ◽  
Dna Sequences ◽  
Fluctuation Analysis ◽  
Chromosome Stability ◽  
Chromosome Loss ◽  
Division Iii ◽  
Daughter Cells ◽  
Chromosome Iii ◽  
Linear Chromosomes

The linear chromosomes of eukaryotes contain specialized structures to ensure their faithful replication and segregation to daughter cells. Two of these structures, centromeres and telomeres, are limited, respectively, to one and two copies per chromosome. It is possible that the proteins that interact with centromere and telomere DNA sequences are present in limiting amounts and could be competed away from the chromosomal copies of these elements by additional copies introduced on plasmids. We have introduced excess centromeres and telomeres into Saccharomyces cerevisiae and quantitated their effects on the rates of loss of chromosome III and chromosome VII by fluctuation analysis. We show that (i) 600 new telomeres have no effect on chromosome loss; (ii) an average of 25 extra centromere DNA sequences increase the rate of chromosome III loss from 0.4 x 10(-4) events per cell division to 1.3 x 10(-3) events per cell division; (iii) centromere DNA (CEN) sequences on circular vectors destabilize chromosomes more effectively than do CEN sequences on 15-kb linear vectors, and transcribed CEN sequences have no effect on chromosome stability. We discuss the different effects of extra centromere and telomere DNA sequences on chromosome stability in terms of how the cell recognizes these two chromosomal structures.

scholarly journals Effects of excess centromeres and excess telomeres on chromosome loss rates.

1991 ◽  
Vol 11 (6) ◽  
pp. 2919-2928 ◽  
Author(s):  
K W Runge ◽  
R J Wellinger ◽  
V A Zakian
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Division ◽  
Dna Sequences ◽  
Fluctuation Analysis ◽  
Chromosome Stability ◽  
Chromosome Loss ◽  
Division Iii ◽  
Daughter Cells ◽  
Chromosome Iii ◽  
Linear Chromosomes

The linear chromosomes of eukaryotes contain specialized structures to ensure their faithful replication and segregation to daughter cells. Two of these structures, centromeres and telomeres, are limited, respectively, to one and two copies per chromosome. It is possible that the proteins that interact with centromere and telomere DNA sequences are present in limiting amounts and could be competed away from the chromosomal copies of these elements by additional copies introduced on plasmids. We have introduced excess centromeres and telomeres into Saccharomyces cerevisiae and quantitated their effects on the rates of loss of chromosome III and chromosome VII by fluctuation analysis. We show that (i) 600 new telomeres have no effect on chromosome loss; (ii) an average of 25 extra centromere DNA sequences increase the rate of chromosome III loss from 0.4 x 10(-4) events per cell division to 1.3 x 10(-3) events per cell division; (iii) centromere DNA (CEN) sequences on circular vectors destabilize chromosomes more effectively than do CEN sequences on 15-kb linear vectors, and transcribed CEN sequences have no effect on chromosome stability. We discuss the different effects of extra centromere and telomere DNA sequences on chromosome stability in terms of how the cell recognizes these two chromosomal structures.

scholarly journals Acquisition and processing of a conditional dicentric chromosome in Saccharomyces cerevisiae

1989 ◽  
Vol 9 (3) ◽  
pp. 1368-1370
Author(s):  
A Hill ◽  
K Bloom
Keyword(s):  
Saccharomyces Cerevisiae ◽  
High Frequency ◽  
Dicentric Chromosome ◽  
Dicentric Chromosomes ◽  
Chromosome Iii ◽  
Derivatives Of ◽  
In Cells

The introduction of a conditional centromere into chromosome III of Saccharomyces cerevisiae provided an opportunity to evaluate phenotypic and karyotypic consequences in cells harboring dicentric chromosomes upon entry into mitosis. A mitotic pause ensued, and monocentric derivatives of chromosome III were generated at a high frequency.

scholarly journals Acquisition and processing of a conditional dicentric chromosome in Saccharomyces cerevisiae.

1989 ◽  
Vol 9 (3) ◽  
pp. 1368-1370 ◽  
Author(s):  
A Hill ◽  
K Bloom
Keyword(s):  
Saccharomyces Cerevisiae ◽  
High Frequency ◽  
Dicentric Chromosome ◽  
Dicentric Chromosomes ◽  
Chromosome Iii ◽  
Derivatives Of ◽  
In Cells

The introduction of a conditional centromere into chromosome III of Saccharomyces cerevisiae provided an opportunity to evaluate phenotypic and karyotypic consequences in cells harboring dicentric chromosomes upon entry into mitosis. A mitotic pause ensued, and monocentric derivatives of chromosome III were generated at a high frequency.

scholarly journals INDUCTION OF INTRACHROMOSOMAL RECOMBINATION IN YEAST BY INHIBITION OF THYMIDYLATE BIOSYNTHESIS

Genetics ◽  
1986 ◽  
Vol 114 (2) ◽  
pp. 375-392
Author(s):  
B A Kunz ◽  
G R Taylor ◽  
R H Haynes
Keyword(s):  
Gene Conversion ◽  
Dna Sequences ◽  
Dna Hybridization ◽  
Crossing Over ◽  
Repeated Dna ◽  
Yeast Saccharomyces Cerevisiae ◽  
Haploid Strain ◽  
Reciprocal Exchange ◽  
Coli Plasmid ◽  
Antifolate Drugs

ABSTRACT The biosynthesis of thymidylate in the yeast Saccharomyces cerevisiae can be inhibited by antifolate drugs. We have found that antifolate treatment enhances the formation of leucine prototrophs in a haploid strain of yeast carrying, on the same chromosome, two different mutant leu2 alleles separated by Escherichia coli plasmid sequences. That this effect is a consequence of thymine nucleotide depletion was verified by the finding that provision of exogenous thymidylate eliminates the increased production of Leu+ colonies. DNA hybridization analysis revealed that recombination, including reciprocal exchange, gene conversion and unequal sister-chromatid crossing over, between the duplicated genes gave rise to the induced Leu+ segregants. Although gene conversion unaccompanied by crossing over was responsible for the major fraction of leucine prototrophs, events involving reciprocal exchange exhibited the largest increase in frequency. These data show that recombination is induced between directly repeated DNA sequences under conditions of thymine nucleotide depletion. In addition, the results of this and previous studies are consistent with the possibility that inhibition of thymidylate biosynthesis in yeast may create a metabolic condition that provokes all forms of mitotic recombination.

Highly repeated DNA sequences in birds: The structure and evolution of an abundant, tandemly repeated 190-bp DNA fragment in parrots

Genomics ◽  
1992 ◽  
Vol 14 (2) ◽  
pp. 462-469 ◽  
Author(s):  
Cort S. Madsen ◽  
Dineke H. de Kloet ◽  
Jean E. Brooks ◽  
Siwo R. de Kloet
Keyword(s):  
Dna Sequences ◽  
Repeated Dna ◽  
Repeated Dna Sequences ◽  
Highly Repeated Dna ◽  
Dna Fragment
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